Objective To establish a malignant transformed human fetal osteoblastic human cells from the immortalized cell line hFOB1.19 in order to explore the molecular mechanism in tumorigenesis of osteosarcoma. Methods hFOB1.19 cells were treated sequentially by an initiated factor, N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) and a promoter, 12-O-tetradecanoyl phorbol-13-acetate (TPA). The features of malignancy of transformed cells were identified by cell morphology, DNA content analysis, colony forming frequency on soft agar and tumorigenetic test in nude mice. Results Continuous passaging after the treatments resulted in the formation of a few paramorph foci and exhibited in an extensively random orientation. The poly-ploid of DNA was 81.08% in experiment group, much higher than that in control group (55.03%). Compared with that of negative control cell, colony formation efficiency of transformed cells in semisolid agar showed a a significant increase. The transformed cells formed tumors subcutaneously in the nude mice, which were verified to be poorly differentiated osteosarcoma histopathological examination. Conclusion Mocking the process of malignant transformation of human cells, we establish malignant transformed immortalized human fetal osteoblastic human cells (hFOB1.19) model.

OBEJECTIVE Gecko has been clinically used in China for many years. It has been proved that the gecko polypeptide mixture(GPM)extracted from gecko could inhibit the growth of multiple types of tumor cells.In order to investigate the possible anti-tumor molecular mechanisms of GPM,we used RNA-seq technology to identify the differentially expressed genes of human hepatocellular carci-noma(HCC)HepG2 cells treated with or without GPM.METHODS The HepG2 cells were treated with different concentration of GPM(0,0.1,0.2,0.3,0.4 mg·mL-1)for 6 h,12 h and 24 h,respectively.MTT assay was used to detect the viability of HepG2 cells. DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells.Western blot analysis was applied to observe the expres-sion of apoptosis- related proteins and endoplasmic reticulum stress (ERs)-related proteins in HepG2 cells.Flow cytometry was also applied to detect reactive oxygen species(ROS)generation.In this report, we showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner.We applied many analysis methods,including differentially expressed genes analysis,Gene Ontology(GO)enrichment analysis,KEGG pathway enrichment analysis,protein-protein interaction network analysis to screen out possible molecular mechanisms.RESULTS ER-nucleus signaling pathway, cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM.GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway. The GPM could induce ROS generation and up-regulate ERs-related proteins. CONCLUSION The present study revealed the potential anti-tumor mechanism of GPM.

Objective To investigate the effect of fasudil on vascular endothelial function in patients with coronary slow flow( CSF) . Methods Eighty?two patients with CSF and normal coronary angiography were selected and randomly divided into conventional treatment group and fasudil group, 41 cases in each group. Patients in conventional treatment group were given conventional treatment( aspirin,nitrates and atorvasta?tin) ,while patients in the fasudil group were given fasudil on the basis of conventional treatment. The angina pectoris,TIMI,endothelial?dependent flow?mediated vasodilation( FMD) ,the levels of plasma nitric oxide( NO) , endothelin?1( ET?1) and Rho kinase( ROCKI) of the brachial artery were observed in the two groups before and after two weeks of treatment. Results The total effective rate of fasudil group was 87. 80%,higher than that of conventional treatment group of 65. 85%,the difference was significant(χ2=68. 176,P<0. 05) . TIMI,FMD im?proved in the fasudil group after treatment compared with before treatment, the difference was significant ( t =4. 37,4. 43;P<0. 05);plasma NO level increased compared with before treatment(t=5. 63,P<0. 01),while ROCKI,ET?1 level decreased(t=6. 19,5. 66;P<0. 01). Plasma NO,ET?1,ROCKI and FMD,TIMI of conven?tional treatment had no significantly changes before and after treatment(P<0. 05). The post?treatment of NO, FMD,TIMI levels in fasudil group were significantly increased compared with conventional group ( ( 36. 17 ±7. 64) μmol/L vs. (24. 99±8. 96) μmol/L,(9. 96±1. 76)% vs. (5. 86±1. 45)%,17. 53±5. 81 vs. 29. 71 ±7. 83;t=4. 06,4. 18,5. 41;P<0. 05),while ROCKI,ET?1 levels in fasudil group were significantly decreased compared with conventional group((19. 57±1. 33) μg/L vs. (34. 38±1. 51) μg/L,(14. 36±6. 05) ng/L vs. (20. 95±6. 57) ng/L;t=3. 87,4. 36,P<0. 01). Conclusion Fasudil can significantly improve the vascular en?dothelial function in patients with CSF.

To investigate the structure disruption of BSA (1mg/ml, dissolved in PBS) induced by ultrasonication and the French press. The BSA solution was passed through the French press and received ultrasound irradiation, and then detected by HPLC(High-performance liquid chromatography),DLS(Dynamic Light Scattering),CD(Circular Dichroism)and nondenaturing SDS-PAGE. Detection results showed that BSA was polymerized after ultrasound irradiation and the polymerization can be reduced by adding mannitol (free radical scavenger). This means that the free radical play an important role in this process. However, the BSA passing through the French press for several times wasn’t polymerized, and the secondary structure was somewhat destroyed. These results suggested that ultrasound irradiation and French press destroy the molecular structure in different manners, so that the suitable cell lyses methods should be selected according to the characteristics of the protein.

AlM:To observe the relationship between axial length and complications of phacoemulsification with intraocular lens ( lOL) implantation in high axial myopia eyes and normal axis eyes.
METHODS: A retrospective review of 843 consecutive patients ( 1 042 eyes ) of cataract extraction with phacoemulsification and lOL implantation in our hospital from February 2012 to February 2013 was performed. The patients were divided into two groups according to the axial length: 853 eyes were in normal axis group ( 21-24mm) and 189 eyes were in high axial myopia group (≥26mm). The two groups were compared regarding surgical complications, such as vitreous loss, posterior capsular rupture, nucleolus drop, and abnormal location of lOL.
RESULTS:Age was a risk factor in both groups. There was positive correlation between age and surgical complications, and between axial length and surgical complications, especially for complications with posterior capsular rupture and vitreous loss.
CONCLUSlON:As the results illustrate, in this survey, age and high axial lengthare statistically significant risk factors for incidence of complications of phacoemulsification. Anticipation of these complications and also preparation and prophylactic measures may decrease incidence of these complications.

Using a H₂O₂-induced BRL cell senescence model, we investigated the anti-aging effects of drug-containing serums of Erzhi Wan (EZW) and various polar extracts (petroleum ether, ethyl acetate, n-butanol, water, and iridoid glycoside-enriched fractions). Cell viability was detected by MTT assay. Cell senescence was evaluated with β-galactosidase staining assay. Intracellular reactive oxygen species (ROS) were measured by flow cytometry. UFLC-Q-TOF-MS/MS was used to identify chemical components in EZW and the extracts, and molecular docking technology was used to predict the anti-aging components of EZW. Results showed that treatment of cells with 600 μmol·L^-1 H₂O₂ for 72 h markedly induced cell senescence, inhibited cell proliferation and increased intracellular β-galactosidase activity and ROS levels. If cells were pretreated with drug-containing serum of EZW this induction of senescence was decreased. A total of 49 chemical compounds were identified in EZW by liquid chromatography-mass spectrometry, 14 of these were identified by molecular docking as potential active ingredients. Based on these analyses, and the in vitro experiments with polar extracts, we conclude that the anti-aging components of EZW are triterpenes, flavonoids and phenyl alcohols, providing a basis for further elucidation of the active agents and mechanism of the anti-aging effect of EZW.

The present study aims to investigate the lignan constituents from Sambucus williamsii and their proliferation effects on osteoblast-like UMR106 cells. Seven compounds were isolated and purified by macroporous resin D101, silica gel, Sephadex LH-20, Toyopearl HW-40, ODS column chromatographies and Preparative HPLC(C-18). Their structures were elucidated by spectroscopic methods as threo-guaiacylglycerol-beta-0-4'-conifery ether (1), lirioresinol A (2), 1-hydroxypinoresinol (3), 5-methoxybalanophonin (4), balanophonin (5), 5-methoxy-trans-dihydrodehydrodiconiferyl alcohol (6), and p-hydroxybenzaldehyde (7). Compounds 3-7 were obtained from this genus for the first time. The proliferation effects of all isolated compounds on osteoblast-like UMR106 cells were determined. Compounds 1-7 (1 x 10(-12)-1 x 10(-7) mol x L(-1)) increased UMR106 cell proliferation to some extent.
Cell Line ; Cell Proliferation ; drug effects ; Lignans ; isolation & purification ; pharmacology ; Osteoblasts ; cytology ; drug effects ; Plant Stems ; chemistry ; Sambucus ; chemistry

A in vitro platelet aggregation bioassay was developed for the quality control of XST capsules. The in vitro anti-platelet aggregation effect in rats was observed to detect the bioactivity of XST capsules. Panax notoginseng saponins and Xuesaitong lyophilizedpowder for injection were taken as standard control substances to determine the potency. According to the results, XST capsules showeda significant inhibitory effect on thrombin-induced platelet aggregation in a dose-dependent manner. The in vitro anti-platelet activity oflyophilized powder for injection was stabler than that of Panax notoginseng saponins, and so suitable to serve as a standard control substance. The biological potency of XST capsules compared with standard control substance was detected by using parallel line assay. According to the results, the established bioassay method had a good repeatability (RSD 2.92%). The sample test results could pass thereliability test(linear deviation P > 0.05, parallel deviation P > 0.05). This bioassay method could be used as one of the complementary quality control methods for XST capsules.
Animals ; Capsules ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Male ; Panax notoginseng ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology

OBJECTIVETo observe the changes of clinical conditions, and measure the GCF volume during the experimental gingivitis in Chinese for studying the relationship between the GCF volume and the development of gingival inflammation.

METHODS11 young male subjects with healthy gingiva, who had no systemic diseases, were selected for this study. GCF samples (18 teeth/person) were collected with strips of filter paper and the clinical parameters were recorded at the baseline (0 day), the 7th, 14th, 21st day (without oral hygiene), and 28th day (7 days after reestablishing oral hygiene) during experimental gingivitis. The GCF volume was measured by weighting.

RESULTSDuring the experimental gingivitis, all of the clinical parameters plaque index (PLI), bleeding index (BI), gingival index (GI) and probing index (PD) increased gradually following the plaque assembling, there were significant differences comparing the data of baseline with the data of afterwards without oral hygiene. On the 28th day, the data reduced to the level of baseline rapidly. The amount of GCF had the same tendency with the clinical parameters during the experimental gingivitis. There was positive correlation between the amount of GCF and clinical parameters.

CONCLUSIONThe amount of GCF can reflect the development of gingival inflammation.

Adult ; Dental Plaque ; complications ; Diet ; Gingival Crevicular Fluid ; physiology ; Gingivitis ; etiology ; Humans ; Male

OBJECTIVETo measure the permeability of human temporomandibular joint (TMJ) disc and cartilage to provide basic parameter for oral biomechanics and tissue engineering, and analyze its mechanisms of pathology and load-release.

METHODSConfined compression method was used to measure the permeability (k value) of four cadavers' TMJs, which were sampled into three parts: disc, condyle and glenoid fossa with different diameters (2 mm, 3 mm and 4 mm). All 128 samples were tested with correspond diameter indenter.

RESULTSLarger the sample diameter was, higher the k value became. The highest k value appeared in the disc while the lowest appeared in glenoid fossa.

CONCLUSIONIn normal condition, TMJ can suffer huge load by decreasing its permeability. Disc is weakest for the higher permeability, it's easy-damaged region is an initiated factor of TMJ disease.

Cartilage ; Humans ; Permeability ; Temporomandibular Joint Disc ; Tissue Engineering